Demethylbelamcandaquinone B from Marantodes pumilum var. alata (Blume) Kuntze inhibits osteoclast differentiation in RAW264.7 cells

Objective: To investigate the bone-resorbing effect of demethylbelamcandaquinone B (Dmcq B) extracted from Marantodes pumilum var. alata on osteoclast differentiation in RAW264.7 cells. Methods: RAW264.7 macrophages were differentiated using RANKL into osteoclast-like cells. Then, they were treated with 10 μg/mL Marantodes pumilum var. alata crude aqueous extract, 5 μg/ mL dichloromethane fraction, and 0.6 μg/mL Dmcq B and 0.06 μg/ mL estradiol. Tartrate-resistant acid phosphatase 5b (TRACP 5b) as an osteoclast phenotypic marker was determined by TRACP staining and TRACP 5b colometric assay, and bone-resorbing pits were examined. The gene expression of pro-inflammatory cytokines (TNF-α and IL-6) was measured. Moreover, the protein expressions of pro-inflammatory cytokines (TNF-α and IL-6) and estrogen receptors were evaluated. Results: Marantodes pumilum var. alata crude aqueous extract and Dmcq B inhibited RANKL-stimulated osteoclast differentiation as evidenced by size reduction of giant multinucleated osteoclast cells, decreased TRACP 5b activity as well as the subsiding of resorbed pit area compared with normal control. In addition, they reduced the gene and protein expressions of TNF-α and IL-6. Marantodes pumilum var. alata, Dmcq B, and estradiol treatments increased the protein expressions of estrogen receptors alpha and beta in osteoclasts. Conclusions: Marantodes pumilum var. alata and its active compound, Dmcq B can inhibit osteoclast differentiation.

Rol de los fitoestrógenos en la calcificación vascular e interacciones óseo-vasculares / Role of phytoestrogens in vascular calcification and bonevascular interactions

La osteoporosis y las enfermedades cardiovasculares son patologías prevalentes en mujeres posmenopáusicas. La calcificación vascular es un proceso en el que se produce una distorsión de la arquitectura natural del tejido arterial con una transformación símil osteogénica. La fisiología vascular y la osteogénesis (formación y remodelación ósea) comparten una complejidad metabólica y funcional crítica, que ha sido poco explorada en forma conjunta, lo que ha impulsado la concepción del Eje Óseo-Vascular como nueva área de investigación, con una visión de estudio integradora con la finalidad de identificar vínculos entre ambos sistemas. En virtud de la controversia planteada sobre los riesgos/beneficios de la terapia de reemplazo hormonal para prevenir enfermedades asociadas a la menopausia, se ha incentivado la búsqueda de nuevas opciones de tratamiento. Los fitoestrógenos, como compuestos nutracéuticos, surgen como una potencial alternativa terapéutica. En particular, las isoflavonas presentan gran analogía estructural con el estrógeno humano 17ß-estradiol, lo que les permite unirse al receptor de estrógenos e inducir acciones estrogénicas tanto en células animales como humanas. Basado en la experiencia propia como en lo reportado en la bibliografía, este artículo analiza la información disponible sobre las acciones vasculares y óseas de los fitoestrógenos (específicamente la isoflavona genisteína), con una visión de ciencia traslacional. Es de esperar que los avances en el conocimiento derivado de la ciencia básica, en un futuro cercano, pueda contribuir a decisiones clínicas a favor de promover terapias naturales de potencial acción dual, para la prevención de enfermedades de alta prevalencia y significativo costo social y económico para la población. (AU)

Osteoporosis and cardiovascular diseases are prevalent diseases in postmenopausal women. Vascular calcification is a cellmediated process that leads to the loss of the natural architecture of the arterial vessels due to osteogenic transdifferentiation of smooth muscle cells, and matrix mineralization. Vascular physiology and osteogenesis (bone formation and remodeling) share a critical metabolic and functional complexity. Given the emerging integrative nature of the bonevascular axis, links between both systems are a matter of ongoing interest. In view of the controversy stated about the risks/benefits of hormone replacement therapy to prevent diseases associated with menopause, phytoestrogens arise as a potential natural therapeutic alternative. In particular, isoflavones have a strong structural analogy with the human estrogen 17ß-estradiol, that allows them to bind to the estrogen receptor and induce estrogenic actions in animal and human cells. Based in on our own experience and the information available in the literature, in this paper we provide an overview of the role of phytoestrogens on vascular and bone tissues, with focus on Genistein actions. We wish that the basic knowledge acquired may contribute to guide clinical decisions for the promotion of natural therapies for the treatment of diseases that conspire against human health. (AU)

Effects Of Short And Long-Term Administration Of Alfalfa On Testicular Histomorphometry In Rats

Alfalfa (Medicago sativa) is a plant with phytoestrogenic properties, which has been used as a major part of diets in husbandry. Since there are controversial reports related to the effects of alfalfa consumption on animal fertility, its effects on rat testicular tissue were assessed in the present study. Control (n=15) and alfalfa (n=15) groups were fed with ordinary rat chow and ordinary rat chow plus alfalfa, respectively. Testicles were removed after 30, 45, and 60 days of consumption, and tissue sections were prepared to assess histomorphometric changes related to alfalfa consumption.Based on the results, there was no significant difference in length, width, and volume of testes of treated rats to control in all groups. But the number of testicular spermatogonia cells, primary spermatocyte cells, primary spermatid cells, testicular spermatozoid cells and Leydig cells significantly or insignificantly increased in rats that received alfalfa for 30 days but all of these parameters insignificantly decreased in rats that received alfalfa for 60 days. The cause of these changes may be due to estrogenic or anti-estrogenic, antioxidant and endocrine effects of alfalfa. Conclusion:Consumption of alfalfa forshort time had only a transient positive effects on testicular tissues but use of alfalfa for 60 days had little destructive effects on testicular tissue in rats. So longer durations of time could be suggested for further research on the effects of alfalfa on rat’s reproduction index

Studies of dietary phytoestrogen from momordica charantia Linn. Seeds / Estudios de fitoestrógeno en la dieta a partir de semillas de momordica charantia Linn

Phytoestrogens are secondary plant metabolites produced by plants that are similar in structure of estrogen. Human consumption of these compounds has been associated with many health benefits. In this study, we investigated the potential phytoestrogen content of dietary Momordica charantia Linn. (Bitter melon) seeds were collected from the Hyderabad-Karnataka regions of India. A phenolic dietary phytoestrogen has been isolated from the crude ethanol extract of Bitter melonseeds. After preparative HPLC whitish amorphous compound was yielded. The HPLC purified compound is subjected to spectral analysis using IR, NMR and MS. The spectral data revealed that the phenolphthalein a phytoestrogenic molecule is present in the ethanol extract of dietary Bitter melonseeds.

Los fitoestrógenos son metabolitos vegetales secundarios, producidos por plantas que tienen una estructura similar al estrógeno. El consumo humano de estos compuestos se ha asociado con muchos beneficios para la salud. En este estudio, investigamos el contenido potencial de fitoestrógenos de Momordica charantia Linn en la dieta. Se recolectaron semillas (melón amargo) de las regiones de Hyderabad-Karnataka de la India. Se ha aislado un fitoestrógeno dietético fenólico del extracto de etanol crudo de semillas de melón amargo. Después de HPLC preparativa, se obtuvo un compuesto amorfo blanquecino. El compuesto purificado por HPLC se somete a análisis espectral usando IR, NMR y MS. Los datos espectrales revelaron que la fenolftaleína, una molécula fitoestrogénica, está presente en el extracto etanólico de las semillas de melón amargo en la dieta.

Effects of Stilbene Glucoside on the Proliferation and Estrogen Receptor Expression of Human Breast Cancer T-47D Cells / 中国药房

OBJECTIVE：To study the effects of stilbene glucoside （TSG）on the proliferation and estrogen receptor （ER）of human breast cancer T- 47D cells ，and to explore its estrogen-like effect and potential mechanism. METHODS ：Taking ER positive human breast cancer T- 47D cells as subjects ，using β-estradiol（β-E2，1×10－8 mol/L）as positive control ，CCK-8 assay was used to detect the cell proliferation after treated with different concentrations of TSG （1×10－8，1×10－7，1×10－6，1×10－5，1×10－4 mol/L）for 24，48，72 h；the cell proliferation rate was calculated. Western blotting assay and RT-PCR methods were adopted to detect the protein and mRNA expression of ER-α and ER-β in cells after treated with low，medium and high concentrations of TSG （1×10－8， 1×10－6，1×10－4 mol/L）for 48 h. RESULTS ：After treated with different concentrations of TSG for 24，48，72 h，the cell proliferation rate of each administration group at each time point （except for β-E2 group at 48 h）increased significantly ，compared with blank group ；those of TSG groups （1×10－5，1×10－6，1×10－7 mol/L）were significantly higher than β-E2 group（P＜0.05 or P＜0.01）. After treated with low ，medium and high concentrations of TSG for 48 h，protein and mRNA expression of ER-α and ER-β in cells were increased significantly，compared with blank group （P＜0.05 or P＜0.01）；protein expression of ER-β in TSG low concentration group ，mRNA expression of ER-α in TSG groups as well as mRNA expression of ER-β in TSG low and high concentration groups were significantly higher than β-E2 group（P＜0.05 or P＜0.01）. CONCLUSIONS ：TSG can induce the in vitro proliferation of T- 47D cells and exert estrogen-like effects by promoting protein and mRNA expression of ER-α and ER-β， which is stronger than that of β-E2 at a certain concentration.

Current status and future of non-genomic effects of phytoestrogens / 药学学报

Phytoestrogens exhibit various pharmacological estrogen-like effects, such as in the prevention and treatment of osteoporosis, cardiovascular diseases, tumors, etc., but the specific mechanism is still unclear. In recent years, estrogen receptor alpha-mediated rapid non-genomic effects have been identified to play an important role in the pathogenesis of estrogen-related diseases. The research of phytoestrogens exerting pharmacological effects through non-genomic effects has also received increasing attention. This article summarizes the research progress in estrogen receptor alpha-mediated non-genomic effects and analyzes the possible involvement of rapid non-genomic effects in certain pharmacological effects of phytoestrogens. The future prospects of estrogen receptor-mediated non-genomic effects by phytoestrogens are also discussed.

Screening of antioxidant components in Scutellariae Radix through mediating Nrf2-ARE signaling pathway based on estrogen receptor / 中国药理学通报

Aim: To establish ARE dual-luciferase reporter assay system and used it to identify the antioxidant substance of Scutellaria baicalensis Georgi. Methods: 293T cells were transiently co-transfected with ARE luciferase reporter plasmid PGL 4. 37 and sea kidney luciferase reporter plasmid PRL-TK. Three major active ingredients of Scutellaria baicalensis Georgi such as scutellarin, baicalein, baicalin and/or estrogen receptor (ER) specific inhibitor were added to Nrf2-ARE luciferase reporter assay system to detect whether they exerted antioxidant effect through the estrogen receptor affecting the Nrf2-ARE signaling pathway. Results: Baicalin (100 μmol · L-1) could obviously activate Nrf2-ARE pathway in 293T cells, and the induced expression was(1. 56 ±0. 01) times that of blank group (P < 0. 01). After pre-administration of ER specific inhibitor, the induced expression decreased to (1. 02 ±0. 23) times, and the antioxidant effect disappeared. After pre-administration of ER and Nrf2-ARE pathway specific inhibitor respectively, ROS in HaCaT cells injured by UVB significantly increased and and SOD was markedly down-regulated by baicalin. Conclusion Baicalin plays antioxidant activity through mediating Nrf2-ARE signaling pathway based on estrogen receptor.

Molecular mechanism of apoptosis of breast cancer SKBR-3 cells induced by cryptanshinone via G protein coupled estrogen receptor( GPER) mediated pathway / 中国中药杂志

The study aimed to illuminate the role of G protein coupled estrogen receptor( GPER) and its mediated PI3 K/AKT signaling pathway in cryptotanshinone( CPT) induced apoptosis of breast cancer SKBR-3 cells,which is GPER positive and ER negative.The apoptosis rate of SKBR-3 cells was tested by Annexin V-FITC/PI staining and apoptosis effector caspase-3 was determined by Western blot. The key proteins in PI3 K/AKT signaling pathway mediated by GPER were detected by Western blot and immunofluorescence technique. Meanwhile,the agonist G1 and antagonist G15 of GPER and antagonist LY294002 of PI3 K were employed in the test to further clarify the effect of GPER and PI3 K/AKT pathway. The results indicated that the apoptosis rate was increased from 4. 7% to46. 1% and 69. 0% after treatment with 0,5,10 μmol·L~(-1) CPT for 48 h( P<0. 01). The expression of PI3 K,AKT and p-AKT were inhibited( P<0. 05 or P<0. 01),while caspase-3 level increased obviously after treatment with CPT( P<0. 01). Importantly,inhibitory effect of PI3 K/AKT signaling pathway by CPT was further enhanced by G1 and attenuated by G15. LY294002 also induced a further inhibition of expression of AKT and p-AKT. The mean fluorescence intensity of AKT and p-AKT could be decreased by CPT. Furthermore,CPT could downregulate GPER expression in SKBR-3 cells( P<0. 01),which could be inhibited by G1 and enhanced by G15.In conclusion,CPT could induce the apoptosis of ER negative and GPER positive breast cancer SKBR-3 cells and the molecular mechanism is related to its regulatory effect of GPER and its mediated PI3 K/AKT signaling pathway.

Effects of Processed Polygonum multiflorum Containing Serum on the Proliferation and the Expression of ER of Human Breast Cancer T- 47D Cells / 中国药房

OBJECTIVE： To study the effects of processed Polygonum multiflorum containing serum on the proliferation and the expression of estrogen receptor （ER） of human breast cancer T-47D cells， and to investigate its phytoestrogen （PE）-like effect. METHODS： Sexually immature SD rats were randomly divided into estradiol valerate （Ev） group （positive control， 0.12 mg/kg）， processed P. multiflorum low-dose and high-dose groups （0.75， 3 g/kg， by crude drug）， low-dose and high-dose processed P. multiflorum+Ev groups （same dose as single drug group）， with 10 rats in each group. Blank group was given constant volume of water intragastrically， and administration groups were given relevant medicine intragastrically； once day and night， for consecutive 4 days. Two hours after last administration， blank serum and containing serum were prepared. T-47D cells were also randomly divided into blank group， Ev group， low-dose and high-dose processed P. multiflorum groups， low-dose and high-dose processed P. multiflorum+Ev groups， and then were cultured in medium which contained 20％ blank serum or drug containing serum. CCK-8 assay was used to detect proliferation rate （PR）. Western blotting assay and RT-PCR were used to detect the protein and mRNA expression of ER-α and ER-β. RESULTS： Compared with blank group， PR of administration groups [each administration group （24 h）， other administration groups （48， 72 h） except for high-dose processed P. multiflorum+Ev group] were increased significantly； high-dose processed P. multiflorum group （72 h） was significantly higher than Ev group， and low-dose processed P. multiflorum+Ev group （72 h） was significantly higher than the same-dose processed P. multiflorum group； high-dose processed P. multiflorum+Ev group （72 h） was significantly lower than the same-dose processed P. multiflorum group （P＜0.05 or P＜0.01）. Relative protein expression of ER-α in Ev group， high-dose processed P. multiflorum group and low-dose processed P. multiflorum+Ev group， relative mRNA expression of ER-α and protein expression of ER-β in administration groups， relative mRNA expression of ER-β in Ev group， low-dose processed P. multiflorum group and processed P. multiflorum+Ev groups were all increased significantly. Relative protein and mRNA expression of ER-α in Ev group were significantly higher than processed P. multiflorum groups and combination groups. Relative protein and mRNA expression of ER-β in Ev group were significantly lower than low-dose processed P. multiflorum+Ev group， but relative mRNA expression of ER-β was significantly higher than processed P. multiflorum groups and high-dose processed P. multiflorum+Ev group. Relative protein and mRNA expression of ER-α and ER-β in low-dose processed P. multiflorum+Ev group as well as relative mRNA expression of ER-β in high-dose processed P. multiflorum+Ev group were significantly higher than the same-dose processed P. multiflorum group. Relative protein and mRNA expression of ER-α in high-dose processed P. multiflorum+Ev group were significantly lower than the same-dose processed P. multiflorum group （P＜0.05 or P＜0.01）. CONCLUSIONS： The processed P. multiflorum containing serum can promote the proliferation of human breast cancer T-47D cells， and play the PE-like role through promoting protein and mRNA expression of ER-α and ER-β. However， the above effects are weaker than estrogen， and the combination of the two may antagonize the effect of estrogen.

Impact of Hydroalcoholic Extract of Humulus Lupulus L. on Sperm Quality, Reproductive Organs and Hormones in Male Rats / 中国结合医学杂志

OBJECTIVE@#To investigate the impact of Humulus Lupulus L. hydroalcoholic extract on the body weights, reproductive organs, sperm quality and hormone levels in male rats.@*METHODS@#By simple random sampling method, seventy male Sprague-Dawley rats were randomly assigned to 7 groups including control group [distilled water, 1 mL/(kg•d)], Tween 80 group [25% Tween 80 solution, 1 mL/(kg•d)], olive oil group [olive oil, 1 mL/(kg•d)], diethyl stilbestrol (DES) group [DES, 100 μg/(kg•body weight)], H50, H150 and H450 [50, 150 and 400 mg/(kg•d) of Humulus Lupulus L extract, respectively]. The administration was performed via gavage once daily for 7 weeks. Body and reproductive organs weights including testes, seminal vesicles, epididymis and prostate were weighted and epididymal sperm quality were determined by digital balance. Blood samples were collected and serum free testosterone (T), luteinizing hormone (LH), follicle-stimulating hormone (FSH), and estrogen (E2) levels were measured by rat specific enzyme-linked immunosorbent assay.@*RESULTS@#The percentage increase in mean body weights of rats in the DES and H50, H150 and H450 groups decreased significantly compared to olive oil and Tween 80 groups (all P0.05).@*CONCLUSION@#Humulus Lupulus L. extract significantly increased the seminal vesicle and testes weights and reduced the sperm motility.

Trifolium pratense: Friable calli, cell culture protocol and isoflavones content in wild plants, in vitro and cell cultures analyzed by UPLC

Abstract Trifolium pratense L., Fabaceae, is a rich source of isoflavones and has become the focus of several studies related to its phytoestrogenic activity. The aim of this study was to establish germination and cell cultures protocol for T. pratense and quantify isoflavones content in cell cultures, in vitro cultured and wild plants harvested in two different seasons. Murashige Skoog medium supplemented with naphthalene acetic acid and kinetin was able to produce the highest formation of friable calli. Calli cultures were analyzed qualitatively after 60 days of culture, and in vitro plants after 30, 45 and 60 days of cultivation. The chemical analysis was performed by ultra performance liquid chromatography, using the linearity curves of daidzein, genistein, formononetin and biochanin A as standards. The concentrations of isoflavones detected in wild plants were different in the two harvest periods and contrasted in content when compared to the in vitro plants. Cell cultures exhibited diverse profiles and concentration of isoflavones, none of which presented the isoflavonoid biochanin A. Pectinase was used to promote reduction of clumps and ended up altering the characteristics of secondary metabolites production in some cultures. Formononetin showed higher concentration in wild red clover samples (15.407 mg g-1), and in the in vitro grown plants the highest concentration was daidzein (17.591 mg g-1) at 60 days. The methods used for this research were effective, and the red clover plants of the analyzed variety can be cultivated in vitro aiming the commercial productivity by having contents greater than or equal to the wild plants in the periods studied, even without the use of elicitors during the cultivation.

Distribution of phytoestrogenic diarylheptanoids and sesquiterpenoids components in Curcuma comosa rhizomes and its related species

Abstract Curcuma comosa Roxb., Zingiberaceae, a phytoestrogen-producing herb with vernacularly named "Wan Chak Mod Loog" in Thailand, has been traditionally used for treatment of gynecologic diseases and sold as food supplement in the market. However, similar rhizomes of its related species may lead to the confusion in the uses of this plant. This study was aimed to investigate the phytochemical constituents of different Curcuma spp. that used as "Wan Chak Mod Loog". Characteristic major compounds were isolated and identified. Phytochemical analysis of 45 Curcuma samples representing Curcuma sp., C. latifolia, and C. comosa were analyzed and compared with their phylogenetic relationship inferred by Amplified Fragment Length Polymorphism analysis. Phytoestrogen diarylheptanoids were found in all samples of C. comosa while sesquiterpenoids including hepatoxic zederone were found in C. latifolia and Curcuma sp. samples.

Research progress in phytoestrogens of traditional Chinese medicine / 中国中药杂志

Phytoestrogens are plant-derived compounds, which have a similarity in structure with human endogenous estrogen 17-β-estradiol. Structural likeness enables phytoestrogens to interact with estrogen receptors, not simply mimicking the effects of human steroidal estrogen but also exhibiting similar and divergent actions. The global literature relating to phytoestrogen in recent years was systematically summarized in this paper. Chemical compositions of phytoestrogens were mainly flavonoids, coumarins, lignans, terpenoids, steroids, etc., with a character of prevention and treatment of perimenopausal syndrome, osteoporosis, cardiovascular disease, metabolic diseases, cancer, regulation of brain function and other pharmacological effects. The mechanisms of action mainly included classical estrogen receptor pathway, epigenetic effect, activation of 5'-adenosyl-phospho-activated protein kinase, inhibition of kinase, activation of peroxisome proliferator-activated receptor, regulation of apoptosis-related proteins, inhibition of nuclear factor κB signaling pathway and so on. According to their efficacy classification, phytoestrogens were mainly distributed in the tonifying medicines, blood-activating and stasis-resolving medicines and heat-clearing medicines. The classical prescriptions with estrogen activity included tonifying formula, Qi-regulating formula and harmonizing formula, etc. This review was aimed at providing a certain reference for the further study of phytoestrogens by researchers and clinicians.

Bidirectional regulation of angiogenesis by phytoestrogens through estrogen receptor-mediated signaling networks / 中国天然药物

Sex hormone estrogen is one of the most active intrinsic angiogenesis regulators; its therapeutic use has been limited due to its carcinogenic potential. Plant-derived phytoestrogens are attractive alternatives, but reports on their angiogenic activities often lack in-depth analysis and sometimes are controversial. Herein, we report a data-mining study with the existing literature, using IPA system to classify and characterize phytoestrogens based on their angiogenic properties and pharmacological consequences. We found that pro-angiogenic phytoestrogens functioned predominantly as cardiovascular protectors whereas anti-angiogenic phytoestrogens played a role in cancer prevention and therapy. This bidirectional regulation were shown to be target-selective and, for the most part, estrogen-receptor-dependent. The transactivation properties of ERα and ERβ by phytoestrogens were examined in the context of angiogenesis-related gene transcription. ERα and ERβ were shown to signal in opposite ways when complexed with the phytoestrogen for bidirectional regulation of angiogenesis. With ERα, phytoestrogen activated or inhibited transcription of some angiogenesis-related genes, resulting in the promotion of angiogenesis, whereas, with ERβ, phytoestrogen regulated transcription of angiogenesis-related genes, resulting in inhibition of angiogenesis. Therefore, the selectivity of phytoestrogen to ERα and ERβ may be critical in the balance of pro- or anti-angiogenesis process.

Research Review on the Estrogen Activity of Traditional Chinese Herbal Medicine / 中国中医药信息杂志

This article sorted out research results of traditional Chinese herbal medicine at home and abroad, cleared up confirmed traditional Chinese herbal medicine with estrogen activity, analyzed the function classifications, and reviewed relevant clinical research progress. It also discussed the concept and connoation of traditional Chinese herbal medicine with estrogen activity, and proposed that the research on traditional Chinese herbal medicine with estrogen activity should clarify whether “bidirectional effect” establishes and the preconditions and requirements of the establishment, with a purpose to solve the security problems about hormone dependency caused by TCM therapy. Meanwhile, the problems about quick-screening experiments, standard establishment, and the transformation of experimental achievements to clinical application are waiting to be solved.

Association between Isoflavones Consumption and Cognitive Function and Comorbidities among Older Adults Residing in the State of Johor, Malaysia

Introduction: This two-phase longitudinal study sought to determine the association between isoflavones intake on cognitive function and comorbidities among older adults from the state of Johor, Malaysia. Methods: Phase I involved baseline data collection to examine the association between isoflavones intake and cognitive function among 400 respondents aged 60-years and above, recruited through multistage random sampling. Phase 2 determined the association between isoflavones intake at the baseline and comorbidities at an 18-month follow-up. The baseline data collected included information on socio-demographics, health status, andunpometric measurements, and dietary intake using a dietary history questionnaire (DHQ). Each participant's cognitive function was evaluated using a mini mental state examination (MMSE), digit span, digit symbol, and geriatric depression scale (GDS). Results: The daily intake of total isoflavones, daidzein, and genistein were 19.1±19.7,11.7±12.3 and 7.6+8.1 mg/day, respectively. There was no significant association between isoflavones intake, and specific cognitive function including global, memory, executive functions, and depression. However, there was a significant association (p 0.05). Conclusion: This study found an association between isoflavones intake and memory function, but not with global cognitive, executive functions, depression, and comorbidities. There is a need to promote adequate isoflavones intake in view of its association with memory function.

Association between Isoflavones Consumption and Cognitive Function and Comorbidities among Older Adults Residing in the State of Johor, Malaysia

Introduction: This two-phase longitudinal study sought to determine the association between isoflavones intake on cognitive function and comorbidities among older adults from the state of Johor, Malaysia. Methods: Phase I involved baseline data collection to examine the association between isoflavones intake and cognitive function among 400 respondents aged 60-years and above, recruited through multistage random sampling. Phase 2 determined the association between isoflavones intake at the baseline and comorbidities at an 18-month follow-up. The baseline data collected included information on socio-demographics, health status, andunpometric measurements, and dietary intake using a dietary history questionnaire (DHQ). Each participant's cognitive function was evaluated using a mini mental state examination (MMSE), digit span, digit symbol, and geriatric depression scale (GDS). Results: The daily intake of total isoflavones, daidzein, and genistein were 19.1±19.7,11.7±12.3 and 7.6+8.1 mg/day, respectively. There was no significant association between isoflavones intake, and specific cognitive function including global, memory, executive functions, and depression. However, there was a significant association (p 0.05). Conclusion: This study found an association between isoflavones intake and memory function, but not with global cognitive, executive functions, depression, and comorbidities. There is a need to promote adequate isoflavones intake in view of its association with memory function.

Effect ofβ-sitosterol on T47D Cell Proliferation and Cell Cycle and Its Mechanisms / 世界科学技术-中医药现代化

This study was aimed to observe the influence of β-sitosterol (BSS) on estrogen receptor (ER) positive the human breast cancer cell line T47D and to study its mechanisms. ER antagonist ICI182 780 was employed to observe the influence on the proliferation. Proliferations of T47D cells influenced by different concentrations of BSS were analyzed by MTT assay. Cell cycle analyses were examined by flow cytometry. The protein expression of cyclin D1 was measured by western blot analysis and cyclin D1 mRNA was quantified by real-time PCR assay. The results showed that BSS in high dose exhibited significant inhibitory effects that were partly antagonized by ICI182 780 and decreased the proliferative index on T47D cells. However, BSS in low dose obviously promoted the proliferation that was completely inhibited by ICI182 780 and increased the proliferative index on T47D cells. The mRNA and protein levels of cyclin D1 were increased in low-dose BSS. The effect was blocked by ICI182 780. It was concluded that BSS in low concentration had phytoestrogenic effect by up-regulating the expression of cyclin D1 via ER pathway.

Evaluation of Estrogenic Effects ofCoicis Semen Aqueous Extracts / 世界科学技术-中医药现代化

This study was aimed to investigate the estrogenic effects ofCoicis Semen in order to preliminarily discuss the mechanism. Mouse uterine weight test and MCF-7 cell proliferation assay were used to evaluate the estrogenic effects ofC. Semen. Reporter gene assays were adopted to explore the action mechanism ofC. Semen. In reporter gene assay, HEK293 cells were co-transfected with pERE-TAL-luc, pβgal-Control, pCXN2-hERα, or pCXN2-hERβ. And the expression of reporter gene luc was controlled by ERE. Mouse uterine weight test showed that compared with the control group, the aqueous extracts ofC. Semen can increase the uterus index of premature female mice (P<0.01). It can significantly promote the proliferation of MCF-7 cells in the medium without estrogen (P<0.01). The reporter gene controlled by ERE technology showed thatwhen mediated by ERα or ERβ respectively, the normalized luciferase activity of aqueous extracts ofC. Semen was significantly higher than activity of the control group (P< 0.05 orP< 0.01). It was concluded that the aqueous extracts ofC.Semen can increase the uterus index of premature female mice and promote the proliferation of MCF-7 cells in the medium without estrogen. We found the estrogenic effects ofC. Semen for the first time. And the estrogenic effects ofC. Semen were mainly mediated by ERβ.

Influence of Genista tinctoria L. or methylparaben on subchronic toxicity of bisphenol A in rats / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To evaluate the influence of an extract of Genista tinctoria L. herba (GT) or methylparaben (MP) on histopathological changes and 2 biomarkers of oxidative stress in rats subchronicly exposed to bisphenol A (BPA).</p><p><b>METHODS</b>Adult female Wistar rats were orally exposed for 90 d to BPA (50 mg/kg), BPA+GT (35 mg isoflavones/kg) or BPA+MP (250 mg/kg). Plasma and tissue samples were taken from liver, kidney, thyroid, uterus, ovary, and mammary gland after 30, 60, and 90 d of exposure respectively. Lipid peroxidation and in vivo hydroxyl radical production were evaluated by histological analysis along with malondialdehyde and 2,3-dihydroxybenzoic acid detection.</p><p><b>RESULTS</b>The severity of histopathological changes in liver and kidneys was lower after GT treatment than after BPA or BPA+MP treatment. A minimal thyroid receptor antagonist effect was only observed after BPA+MP treatment. The abnormal folliculogenesis increased in a time-dependent manner, and the number of corpus luteum decreased. No significant histological alterations were found in the uterus. The mammary gland displayed specific estrogen stimulation changes at all periods. Both MP and GT revealed antioxidant properties reducing lipid peroxidation and BPA-induced hydroxyl radical generation.</p><p><b>CONCLUSION</b>GT L. extract ameliorates the toxic effects of BPA and is proved to have antioxidant potential and antitoxic effect. MP has antioxidant properties, but has either no effect or exacerbates the BPA-induced histopathological changes.</p>